Silicon's Influence on Polyphenol and Flavonoid Profiles in Pea (Pisum sativum L.) under Cadmium Exposure in Hydroponics: A Study of Metabolomics, Extraction Efficacy, and Antimicrobial Properties of Extracts.

The current study aimed to investigate the impact of silicon (Si) supplementation in the form of Na2SiO3 on the metabolome of peas under normal conditions and following exposure to cadmium (Cd) stress. Si is known for its ability to enhance stress tolerance in various plant species, including the mitigation of heavy metal toxicity. Cd, a significant contaminant, poses risks to both human health and the environment. The study focused on analyzing the levels of bioactive compounds in different plant parts, including the shoot, root, and pod, to understand the influence of Si supplementation on their biosynthesis. Metabolomic analysis of pea samples was conducted using a targeted HPLC/MS approach, enabling the identification of 15 metabolites comprising 9 flavonoids and 6 phenolic acids. Among the detected compounds, flavonoids, such as flavon and quercetin, along with phenolic acids, including chlorogenic acid and salicylic acid, were found in significant quantities. The study compared Si supplementation at concentrations of 1 and 2 mM, as well as Cd stress conditions, to evaluate their effects on the metabolomic profile. Additionally, the study explored the extraction efficiency of three different methods: accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and maceration (MAC). The results revealed that SFE was the most efficient method for extracting polyphenolic compounds from the pea samples. Moreover, the study investigated the stability of polyphenolic compounds under different pH conditions, ranging from 4.0 to 6.0, providing insights into the influence of the pH on the extraction and stability of bioactive compounds.

Lipidomics Characterization of the Microbiome in People with Diabetic Foot Infection Using MALDI-TOF MS.

Lipidomic profiling has emerged as a powerful tool for the comprehensive characterization of bacterial species, particularly in the context of clinical diagnostics. Utilizing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), this study aims to elucidate the lipidomic landscapes of bacterial strains isolated from diabetic foot infections (DFI). Our analysis successfully identified a diverse array of lipids in the cellular membranes of both Gram-positive and Gram-negative bacteria, revealing a total of 108 unique fatty acid combinations. Specifically, we identified 26 LPG, 33 LPE, 43 PE, 114 PG, 89 TAG, and 120 CLP in Gram-positive bacteria and 10 LPG, 14 LPE, 124 PE, 37 PG, 13 TAG, and 22 CLP in Gram-negative strains. Key fatty acids, such as palmitic acid, palmitoleic acid, stearic acid, and oleic acid, were prominently featured. Univariate analysis further highlighted distinct lipidomic signatures among the bacterial strains, revealing elevated levels of phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) in Gram-negative bacteria associated with DFI. In contrast, Gram-positive strains demonstrated increased or uniquely fluctuating levels of triglyceride (TAG) and cardiolipin (CLP). These findings not only underscore the utility of MALDI-TOF MS in bacterial lipidomics but also provide valuable insights into the lipidomic adaptations of bacteria in diabetic foot infections, thereby laying the groundwork for future studies aimed at constructing microbial lipid libraries for enhanced bacterial identification.

Metal tolerance and Cd phytoremoval ability in Pisum sativum grown in spiked nutrient solution.

In the presented study, the effects of cadmium (Cd) stress and silicon (Si) supplementation on the pea plant (Pisum sativum L.) were investigated. The tendency to accumulate cadmium in the relevant morphological parts of the plant (roots and shoots respectively)-bioaccumulation, the transfer of this element in the plant (translocation) and the physiological parameters of the plant through indicators of oxidative stress were determined. Model studies were carried out at pH values 6.0 and 5.0 plant growth conditions in the hydroponic cultivation. It was shown that Cd accumulates mostly in plant roots at both pH levels. However, the Cd content is higher in the plants grown at lower pH. The Cd translocation factor was below 1.0, which indicates that the pea is an excluder plant. The contamination of the plant growth environment with Cd causes the increased antioxidant stress by the growing parameters of the total phenolic content (TPC), polyphenol oxidase activity (PPO), the malondialdehyde (MDA) and lipid peroxidation (LP). The results obtained showed that the supplementation with Si reduces these parameters, thus lowering the oxidative stress of the plant. Moreover, supplementation with Si leads to a lower content of Cd in the roots and reduces bioaccumulation of Cd in shoots and roots of pea plants.

Silver Nanoparticle Targets Fabricated Using Chemical Vapor Deposition Method for Differentiation of Bacteria Based on Lipidomic Profiles in Laser Desorption/Ionization Mass Spectrometry.

The global threat of numerous infectious diseases creates a great need to develop new diagnostic methods to facilitate the appropriate prescription of antimicrobial therapy. More recently, the possibility of using bacterial lipidome analysis via laser desorption/ionization mass spectrometry (LDI-MS) as useful diagnostic tool for microbial identification and rapid drug susceptibility has received particular attention because lipids are present in large quantities and can be easily extracted similar to ribosomal proteins. Therefore, the main goal of the study was to evaluate the efficacy of two different LDI techniques-matrix-assisted (MALDI) and surface-assisted (SALDI) approaches-in the classification of the closely related Escherichia coli strains under cefotaxime addition. Bacterial lipids profiles obtained by using the MALDI technique with different matrices as well as silver nanoparticle (AgNP) targets fabricated using the chemical vapor deposition method (CVD) of different AgNP sizes were analyzed by the means of different multivariate statistical methods such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), sparse partial least squares discriminant analysis (sPLS-DA), and orthogonal projections to latent structures discriminant analysis (OPLS-DA). The analysis showed that the MALDI classification of strains was hampered by interference from matrix-derived ions. In contrast, the lipid profiles generated by the SALDI technique had lower background noise and more signals associated with the sample, allowing E. coli to be successfully classified into cefotaxime-resistant and cefotaxime-sensitive strains, regardless of the size of the AgNPs. AgNP substrates obtained using the CVD method were used for the first time for distinguishing closely related bacterial strains based on their lipidomic profiles and demonstrate high potential as a future diagnostic tool for the detection of antibiotic susceptibility.

Isolation and Identification of Lactococcus lactis and Weissella cibaria Strains from Fermented Beetroot and an Investigation of Their Properties as Potential Starter Cultures and Probiotics.

The presence of certain microorganisms in dairy products or silage is highly desirable. Among them are probiotic strains of lactic acid bacteria (LAB), which show many beneficial features, including antimicrobial properties that support the development of beneficial microflora; in addition, owing to their biochemical activity, they influence the nutritional, dietary, and organoleptic properties of food products. Before being placed on the market, each strain requires separate testing to determine its probiotic properties and effectiveness. The aim of this study was to isolate LAB strains from a pickled beetroot sample that could be used in the dairy industry and with the potential to be considered as a probiotic in the future. Two strains identified using the MALDI technique were selected-Lactococcus lactis and Weissella cibaria. The optimal growth conditions of the strains were determined, and their proteolytic properties were assessed with the use of the o-PA reagent and spectrophotometry. The lipid profile was analyzed using the SALDI (surface-assisted laser desorption/ionization) technique and silver nanoparticles. High-performance liquid chromatography was used to assess the ability of the strains to synthesize beneficial metabolites, such as B vitamins (B2, B3, and B9) or lactic acid, and gas chromatography was used to analyze the substances responsible for organoleptic properties. Moreover, the ability to inhibit the growth of pathogenic strains was also tested in the selected strains. Both tested strains demonstrated the desired properties of starter cultures for future use in functional food production, showing that fermented plant products can serve as valuable potential probiotic sources.

Lacticaseibacillus paracasei as a Modulator of Fatty Acid Compositions and Vitamin D3 in Cream.

Butter is an important source of essential fatty acids, lipid-soluble vitamins, and antioxidants in the diet. However, this study showed that the presence of the Lacticaseibacillus paracasei strain has a great influence on the fatty acid profile as well as provitamin D3 and vitamin D3 content in the cream-the raw material from which the butter is obtained. The addition of this lactic acid bacteria enriches the cream in 9-hexadecenoic acid, oleic acid, octadeca-9,12-dienoic acid, and conjugated linoleic acid, which exhibit antimutagenic and anticarcinogenic properties. Moreover, a higher level of monounsaturated fatty acids can extend the shelf life of butter in the future. In the present work, we observed that the presence of lactic acid bacteria contributed to an increase in the level of provitamin D after 6 h of incubation and an increase in the levels of vitamin D3 after 24 and 48 h. Fatty acid profiles and the content of vitamins were largely dependent on the presence of light and mixing, which are probably associated with the status of lipid peroxidation.

Investigation of the mechanism of zearalenone metabolization in different systems: Electrochemical and theoretical approaches.

Mycotoxins are toxic metabolites produced by mold fungi, which commonly contaminate cereal crops. These compounds include zearalenone (ZEA), which may disturb the proper functioning of the endocrine system in mammals. The metabolism of ZEA plays a key role in its toxic properties. The type and amount of produced metabolites may contribute to both the reduction and increase in its pathogenic effect. Therefore, it is extremely important to investigate the possible pathways of zearalenone metabolism. The electrochemical approach may prove alternatives for the in vitro and in vivo methods. For this reason, in this study the electrochemical, theoretical and in vitro assays were applied to determine the possible ZEA metabolism mechanism. Electrochemical processes were conducted in the EC/ESI-MS system. The application of HPLC-MS/MS allowed to characterize the obtained electrochemical products specific for phase I and II. Appropriate conditions for redox identification processes were optimized with regard to such parameters as the potential value, mobile phase and working electrode. In addition, the data obtained by instrumental techniques for ZEA metabolism were compared with those obtained by in vitro methods for HepG2 cell lines. Furthermore, results from quantum mechanical calculations allowed to explain the mechanism and the conformational stability of the reduction products.

Promising Green Technology in Obtaining Functional Plant Preparations: Combined Enzyme-Assisted Supercritical Fluid Extraction of Flavonoids Isolation from Medicago Sativa Leaves.

To elaborate a complete extraction protocol for the enhanced release of biologically active compounds from plant cells, this study aimed to optimize together the parameters of the supercritical fluid extraction (SFE) process (temperature, pressure, and percentage of cosolvent) and enzymatic treatment of plant material (pH, enzyme concentration, time, and temperature) by response surface methodology (RSM). Medicago sativa L. was selected as a plant material due to its richness in phenolics and flavonoids. HPLC-MS/MS analysis allowed evaluating the content of individual bioactive compounds in obtained extracts. The total content of polyphenolic compounds in the extract obtained after two-step optimization was much higher (546 ± 21 µg/g) than in the extract obtained from non-hydrolyzed material (275 ± 23 µg/g) and in the extract obtained by maceration (162 ± 20 µg/g). Furthermore, it was evidenced that extract with the highest content of polyphenolic compounds can support the cellular antioxidant system both as a free radical scavenger and by stimulating the antioxidant enzyme system.

The Study on Molecular Profile Changes of Pathogens via Zinc Nanocomposites Immobilization Approach.

The most critical group of all includes multidrug resistant bacteria that pose a particular threat in hospitals, as they can cause severe and often deadly infections. Modern medicine still faces the difficult task of developing new agents for the effective control of bacterial-based diseases. The targeted administration of nanoparticles can enhance the efficiency of conventional pharmaceutical agents. However, the interpretation of interfaces' interactions between nanoparticles and biological systems still remains a challenge for researchers. In fact, the current research presents a strategy for using ZnO NPs immobilization with ampicillin and tetracycline. Firstly, the study provides the mechanism of the ampicillin and tetracycline binding on the surface of ZnO NPs. Secondly, it examines the effect of non-immobilized ZnO NPs, immobilized with ampicillin (ZnONPs/AMP) and tetracycline (ZnONPs/TET), on the cells' metabolism and morphology, based on the protein and lipid profiles. A sorption kinetics study showed that the antibiotics binding on the surface of ZnONPs depend on their structure. The efficiency of the process was definitely higher in the case of ampicillin. In addition, flow cytometry results showed that immobilized nanoparticles present a different mechanism of action. Moreover, according to the MALDI approach, the antibacterial activity mechanism of the investigated ZnO complexes is mainly based on the destruction of cell membrane integrity by lipids and proteins, which is necessary for proper cell function. Additionally, it was noticed that some of the identified changes indicate the activation of defense mechanisms by cells, leading to a decrease in the permeability of a cell's external barriers or the synthesis of repair proteins.

Lipidomics as a Diagnostic Tool for Prostate Cancer.

The main goal of this study was to explore the phospholipid alterations associated with the development of prostate cancer (PCa) using two imaging methods: matrix-assisted laser desorption ionization with time-of-flight mass spectrometer (MALDI-TOF/MS), and electrospray ionization with triple quadrupole mass spectrometer (ESI-QqQ/MS). For this purpose, samples of PCa tissue (n = 40) were evaluated in comparison to the controls (n = 40). As a result, few classes of compounds, namely phosphatidylcholines (PCs), lysophosphatidylcholines (LPCs), sphingomyelins (SMs), and phosphatidylethanolamines (PEs), were determined. The obtained results were evaluated by univariate (Mann-Whitney U-test) and multivariate statistical analysis (principal component analysis, correlation analysis, volcano plot, artificial neural network, and random forest algorithm), in order to select the most discriminative features and to search for the relationships between the responses of these groups of substances, also in terms of the used analytical technique. Based on previous literature and our results, it can be assumed that PCa is linked with both the synthesis of fatty acids and lipid oxidation. Among the compounds, phospholipids, namely PC 16:0/16:1, PC 16:0/18:2, PC 18:0/22:5, PC 18:1/18:2, PC 18:1/20:0, PC 18:1/20:4, and SM d18:1/24:0, were assigned as metabolites with the best discriminative power for the tested groups. Based on the results, lipidomics can be found as alternative diagnostic tool for CaP diagnosis.

Combination of electrochemical unit and ESI-MS in fragmentation of flavonoids.

INTRODUCTION: Predictive approaches on the activity of natural compounds based on the fragmentation by instrumental techniques are important for consideration of such molecules as drug candidates and defining new structures with promising properties. Since flavonoids are well-known antioxidants, their redox properties can be related to their pharmacological activity. OBJECTIVES: In this work, the potential of electrochemical unit coupled to electrospray ionisation mass spectrometry (ESI-MS) was assessed for fragmentation activity relationships studies of selected flavonoids. METHODOLOGY: Methodology of this research included electrochemical conversion of standards of flavonoids at different pH values and their further analysis with the use of ESI-MS. In addition, signals obtained from the blank samples were also identified and used for interpretation due to electrochemical nature of the ESI source. Half maximal inhibitory concentration (IC50 ) values of flavonoids for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant activity assays were analysed for possible correlation with the structures of flavonoids and products of electrochemical conversion. RESULTS: Fragmentation activity relationships were suggested using the proposed approach and for some of the flavonoids it was not specific enough to determine the input of a particular structural feature to the activity, but for others they were in agreement with those found in the literature. Obtained results showed potential of the proposed approach for application in plant sciences as a fast pre-screening tool for newly isolated bioactive compounds.

Lipidomic analysis of lactic acid bacteria strains by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Analysis by MALDI-TOF mass spectrometry and gas chromatography-mass spectrometry was used to characterize the lipid profile of 3 lactic acid bacteria strains. By gas chromatography coupled with mass spectrometry, 23 fatty acids were identified. Dominant acids were palmitic (C16:0), oleic (C18:1), and α-linoleic acid (C18:3n-3) for Lactobacillus paracasei; for Lactococcus lactis they were palmitic (C16:0), gondoic (C20:1), myristoleic (C14:1), and eicosadienoic acid (C20:2), respectively; and in the case of Lactobacillus curvatus were C18:1, C18:2n-6, and C16:0, respectively. The effect of the medium on fatty acid composition was also determined. In addition, the fatty acid profile was also compared using MALDI MS analysis. The MALDI-TOF MS was used for qualitative analysis and identification of bacterial lipids. Phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine, triacylglycerols, and ceramides were the most abundant species in lactic acid bacteria. One hundred different combinations of fatty acids in polar and nonpolar lipids have been identified, including 11 phospholipids (18 phosphatidylglycerol, 16 phosphatidylethanolamine, 10 phosphatidylinositol, 8 phosphatidylcholine, 4 lyso-phosphatidylethanolamine, 3 lyso-phosphatidylcholine, 3 phosphatidylserine, 1 lyso-phosphatidic acid, 1 lyso-phosphatidylglycerol, 1 lyso-phoshatidylinositol, and 1 phosphatidic acid), 23 triacylglycerols, 9 ceramides, and 2 sphingomyelin. The most abundant fatty acids identified were C16:0, C16:1, C18:0, and C18:3. Obtained lipid profiles allowed to distinguish the tested bacterial strains.

New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS.

A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, cyclitols, and their respective products generated through the electrochemistry (EC) process.

The Influence of Plant Material Enzymatic Hydrolysis and Extraction Conditions on the Polyphenolic Profiles and Antioxidant Activity of Extracts: A Green and Efficient Approach.

The aim of this study was to develop a new comprehensive extraction protocol based on green technology for the enhanced release of polyphenolic compounds from plant cells. In this work, extracts from yerba mate and yellow lupine seed were obtained by using three different extraction techniques: maceration, supercritical fluid extraction with co-solvent (SFE) and enzyme assisted-supercritical fluid extraction with co-solvent (EA-SFE). Several experimental parameters such as time, type of solvent and co-solvent as well as CO2 flow rate were selected to obtain the highest extraction efficiency. The chemical profiles in the obtained extracts and their biological activity were evaluated. HPLC-MS/MS analysis indicated that the level of phenolic compounds in extracts from yerba mate obtained by EA-SFE was approximately five times higher than for maceration and 3.2 times higher than for SFE. In the case of extracts from yellow lupine seed an approximately 5.6-fold increase was observed in comparison with maceration and SFE with 96% MeOH, and 2.9 times for SFE with 96% EtOH. The developed protocol with a mix of enzymes commonly applied in the agricultural industry significantly raises the efficiency of liberation of secondary metabolites.

The Influence of Different Forms of Silver on Selected Pathogenic Bacteria.

The application of silver nanoparticles as an antibacterial agent is becoming more common. Unfortunately, their effect on microorganisms is still not fully understood. Therefore, this paper attempts to investigate the influence of silver ions, biologically synthesized silver nanoparticles and nanoparticles functionalized with antibiotics on molecular bacteria profiles. The initial stage of research was aimed at the mechanism determination involved in antibiotics sorption onto nanoparticles' surface. For this purpose, the kinetics study was performed. Next, the functionalized formulations were characterized by Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS) and a zeta potential study. The results reveal that functionalization is a complex process, but does not significantly affect the stability of biocolloids. Furthermore, the antimicrobial assays, in most cases, have shown no increases in antibacterial activity after nanoparticle functionalization, which suggests that the functionalization process does not always generate the improved antimicrobial effect. Finally, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique was employed to characterize the changes in the molecular profile of bacteria treated with various antibacterial agents. The recorded spectra proved many differences in bacterial lipids and proteins profiles compared to untreated cells. In addition, the statistical analysis of recorded spectra revealed the strain-dependent nature of stress factors on the molecular profile of microorganisms.

Use of Lactobacillus paracasei strain for zearalenone binding and metabolization.

The study investigated the zearalenone (ZEA) neutralization process as a consequence of metabolization and binding process by the probiotic bacterial strain Lactobacillus paracasei using high performance liquid chromatography (HPLC). In order to determine the nature of the binding process the kinetic and spectroscopic approach were used. Moreover, the influence of ZEA on L. paracasei metabolism was examined by the determination of the proteome profile of cells and the profile of volatile compounds (VOCs) produced by bacteria cells. For this purpose the Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (MALDI-TOF MS) and headspace solid-phase microextraction coupled to gas chromatography/mass spectrometry (HS-SPME/GC-MS) techniques were used. The obtained results indicate that in the mechanism of ZEA neutralization both - metabolization/biotransformation and binding/biosorption processes are involved. Furthermore, the biotransformation of ZEA to both α- and ß-ZOL with a predominance of ß-ZOL by lactic acid bacteria strain was recorded. The results suggest that the tested microorganism can be used as a potential detoxification agent for grain and feed.

Mechanistic Chromatographic Column Characterization for the Analysis of Flavonoids Using Quantitative Structure-Retention Relationships Based on Density Functional Theory.

This work aimed to unravel the retention mechanisms of 30 structurally different flavonoids separated on three chromatographic columns: conventional Kinetex C18 (K-C18), Kinetex F5 (K-F5), and IAM.PC.DD2. Interactions between analytes and chromatographic phases governing the retention were analyzed and mechanistically interpreted via quantum chemical descriptors as compared to the typical 'black box' approach. Statistically significant consensus genetic algorithm-partial least squares (GA-PLS) quantitative structure retention relationship (QSRR) models were built and comprehensively validated. Results showed that for the K-C18 column, hydrophobicity and solvent effects were dominating, whereas electrostatic interactions were less pronounced. Similarly, for the K-F5 column, hydrophobicity, dispersion effects, and electrostatic interactions were found to be governing the retention of flavonoids. Conversely, besides hydrophobic forces and dispersion effects, electrostatic interactions were found to be dominating the IAM.PC.DD2 retention mechanism. As such, the developed approach has a great potential for gaining insights into biological activity upon analysis of interactions between analytes and stationary phases imitating molecular targets, giving rise to an exceptional alternative to existing methods lacking exhaustive interpretations.

Study of chromatographic behavior of antibiotic drugs and their metabolites based on quantitative structure-retention relationships with the use of HPLC-DAD.

The separation of eleven antibiotics and ten metabolites were studied using high performance liquid chromatography. The C18-PFP octadecyl with integral PFP, C18-AR octadecyl with integral phenyl, C18-HL octadecyl and phenyl phase were used as stationary phases. Mixtures of acetontrile-0.1 % formic acid in water were investigated as mobile phases. The elution order of the target compounds was similar for all four HPLC columns applied. The best separation was obtained using the column with the pentafluorophenylpropyl chain. In addition, in order to optimize the parameters of retention elution for the column and to predict the conditions for the best separation of the active compounds studied biologically the ChromSword software was used. To obtain reliable information of the physicochemical properties and to estimate the relative biological activity of a group of the studied analytes, the QSRR approach was applied. Molecular descriptors were calculated using the HyperChem software. The study was based on multiple linear regression and the results were presented as quantitative structure-retention relationships equations. The QSRR models were determined using 16 molecular descriptors mainly related to the dipole moment (µ), the solvent accessible surface area (SAS), the van der Waals surface area (VWS), the minimum charge (δmin) as well as the polar surface area (PSA). Moreover, structural descriptors of the target compounds were used to describe their chromatographic retention behavior under the optimized HPLC conditions.

Extraction and Determination of Polar Bioactive Compounds from Alfalfa (Medicago sativa L.) Using Supercritical Techniques.

The aim of this research was to select parameters for supercritical extraction with CO2 of Medicago sativa L., considered as functional food, in quarter-technical plant, providing the highest concentration of bioactive polar constituents and simultaneously maintaining the highest efficiency of the process. For the purpose of optimization, mathematical statistics was used. Qualitative analysis of products was performed with supercritical fluid chromatography (SFC). The SFC analysis revealed a proper separation of flavonoids and phenolics acids for dedicated TFC and TPC optimal parameters. The obtained results have proved that it is a possibility to extract polar compounds with non-polar solvent under higher values of pressure and temperature and to enrich product with desired group of bioactive compounds with proper optimization. The proposed extraction technique allows to obtain on an industrial scale, using an environmentally friendly solvent, a preparation rich in biologically active nutrients that can be implemented in the cosmetics, pharmaceutical and food industries.